Aerva lanata: Roots Extract for the analysis of phytochemicals

Abstract

Numerous common illnesses may be prevented or treated with the help of phytochemicals. There is little doubt that identifying and isolating these phytochemicals would benefit human civilisation. Consequently, this research work explores phytochemicals and performs qualitative and quantitative evaluation of the same. The roots of Aerva lanata were harvested and extracted using the maceration process using solvents including chloroform, ethyl acetate, methanol, and water. Additional qualitative and quantitative research was conducted on the topic. According to the findings, water, methanol, ethyl acetate, and chloroform had respective concentrations of 2.32%, 2.90%, 8.14%, and 3.44%. The sole substance detected in the chloroform extract was tannin. The phenol and tannin tests for ethyl acetate were positive. The phytoconstituents flavonoid, phenol, and tannin were considerably more abundant in the methanolic extract. The aqueous extract ultimately tested positive for tannin & flavonoid. The Aerva lanata extract contains additional classes of phenol and flavonoids in addition to the standard used for comparison, according to the results of TLC for phenol and flavonoid analysis. The methanolic extract of Aerva lanata is estimated to have a total phenolic content of 1.380 mg/100 mg, whilst the ethyl acetate and aqueous extracts had phenol contents of 0.866 mg/100 mg and 0.613 mg/100 mg, respectively. Only the methanolic extract's total flavonoid content, which was found to be 1.280 mg/100 mg, was evaluated. Aerva lanata root has a large Aerva lanata: Roots Extract for the analysis of phytochemicals

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Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1349
Aerva lanata: Roots Extract for the analysis of phytochemicals
Dr. M. Shanmugavadivu1, Dr. Priyanka Bankoti2, Dr. Biren N. Shah3,
Tushar Arun Rode4, Dr Balan Paramasivam5, Abhisek Saha6,
Ashish Kumar Tiwari7, Dr. Nihar Ranjan Kar8, Pydiraju Kondrapu9*
1Associate Professor, Dr. N. G. P. Arts and Science College, Coimbatore-641048, Tamil Nadu,
India
2Professor, Shri guru ram rai university Dehradun
3Professor, Shree Naranjibhai Lalbhai Patel College of Pharmacy, Bardoli Mota Road,
Bardoli, Gujara-394345
4Assistant Professor, P. W. College of Pharmacy, Moha Phata, Dhamangaon Road, Yavatmal.
445001.
5Professor, The Erode College of Pharmacy, Erode -638112, Tamil Nadu.
6Associate Professor in Chemistry, Tufanganj College, West Bengal-736160, India
7Assistant Professor, Krishnarpit Institute of Pharmacy, Iradatganj, Ghoorpur, Prayagraj,
Uttar Pradesh, Pin-212107
8Assistant Professor, Centurion University of Technology and Management, Gopalpur,
Balasore, Odisha, India
9Assistant Professor, Aditya Pharmacy College, Surampalem, India
*Corresponding Author:Pydiraju Kondrapu,
Assistant Professor, Aditya Pharmacy College, Surampalem, India
Email: pydikondrapu604@gmail.com
Article History: Received:10/7/23 Revised: 18/7/23 Accepted: 25/7/23
Abstract:
Numerous common illnesses may be prevented or treated with the help of phytochemicals.
There is little doubt that identifying and isolating these phytochemicals would benefit human
civilisation. Consequently, this research work explores phytochemicals and performs
qualitative and quantitative evaluation of the same. The roots of Aerva lanata were harvested
and extracted using the maceration process using solvents including chloroform, ethyl acetate,
methanol, and water. Additional qualitative and quantitative research was conducted on the
topic. According to the findings, water, methanol, ethyl acetate, and chloroform had respective
concentrations of 2.32%, 2.90%, 8.14%, and 3.44%. The sole substance detected in the
chloroform extract was tannin. The phenol and tannin tests for ethyl acetate were positive. The
phytoconstituents flavonoid, phenol, and tannin were considerably more abundant in the
methanolic extract. The aqueous extract ultimately tested positive for tannin & flavonoid. The
Aerva lanata extract contains additional classes of phenol and flavonoids in addition to the
standard used for comparison, according to the results of TLC for phenol and flavonoid
analysis. The methanolic extract of Aerva lanata is estimated to have a total phenolic content
of 1.380 mg/100 mg, whilst the ethyl acetate and aqueous extracts had phenol contents of 0.866
mg/100 mg and 0.613 mg/100 mg, respectively. Only the methanolic extract's total flavonoid
content, which was found to be 1.280 mg/100 mg, was evaluated. Aerva lanata root has a large

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1350
number of bioactive chemicals, which might be further examined for pharmacological actions,
according to the results that were acquired.
Keywords: Herbal medicines, Aerva lanata, Thin layer chromatography, Total phenol content,
Total flavonoids content, Phytochemicals, Medicinal plant.
DOI: 10.48047/ecb/2023.12.si12.122
Introduction
It is well acknowledged that plants constitute a crucial part of the biodiversity of the earth and
one of its most important natural resources.The history of human civilization can be traced
back to the dawn of the healing arts. Some of the plant's chemical constituents, which have a
clear physiological effect on the human body, have medical significance (Pieroet al.,2012;
Prakash Sharma, 2014).
Primary and secondary metabolites known as phytochemicals are found naturally in many areas
of plants and act as a plant's defense mechanism against numerous pathogens. Primary
metabolites (carbohydrates, lipids, and proteins) are directly involved in plant development and
mechanism. Secondary metabolites, such as alkaloids, phenolics, sterols, steroids, essential
oils, lignins, and tannins, among others, are thought of as the end products of primary
metabolites and are engaged in metabolic activity (Ali and Alqurainy, 2006; Velu et al., 2018;
Frisvadet al., 2007). Phytochemicals are naturally occurring chemical substances that are
physiologically active and are present in plants. They shield plant cells from environmental
dangers such pollution, stress, dehydration, UV exposure, and pathogenic attack. These
substances, also referred to as secondary plant metabolites, are advantageous for human health.
They are believed to work as synergistic agents, enabling the body to utilize nutrients more
effectively. Low toxicity, low cost, easy accessibility, and biological properties like antioxidant
activities, antimicrobial effects, modulation of detoxification enzymes, stimulation of the
immune system, reduction of platelet aggregation, modulation of hormone metabolism, and
antineoplastic properties are some of the advantages of phytochemicals.Phytochemicals have
vital properties to prevent or treat various common diseases, even though they are not necessary
nutrients or needed by the human body to sustain life. Numerous studies have been conducted
to demonstrate the health advantages of phytochemicals as a result of this feature (Mendoza &
silva, 2018; Nyamai et al., 2016).
The plant Aerva lanata (Linn) Juss. ex. Schult is widely used in urinary disorders in southern
part of India as a source of Pashanabheda. It is commonly known as Gorakha ganja a member
of Amaranthaceae, usually found as weed on mountains and bare ground. It is an herb which
trails on the ground with many branches and leaves are alternately arranged with fine hairs
above and with wooly beneath. Flowers are greenish white in clusters. Since years many
researches have been carried out to elicit the diuretic & anti-urolithic activity of this plant.
Besides, it has been proven for many more pharmacological activities like anti-diarrhoeal, anti-
hyperglycameic, anti-oxidant, anti-helmentic, and analgesic. In addition, various phyto
chemical investigations reveal the presence of steroids, tannins, flavonoids, nutrients,
terpenoids in different parts of the plant (Nagaratnaet al., 2014; Bitastaand Madan, 2016;
Athiraand Nair, 2017). Thus, this study paper deals with exploring the phytochemicals &
performing qualitative & quantitative estimation of the same.

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1351
Material & Methods
Collection of plant material
Roots of Aerva lanata Linn were collected from local area of Bhopal in month of January,
2022. Drying of fresh plant parts was carried out in sun but under the shade.
Extraction procedure
Following procedure was adopted for the preparation of extract from the shade dried and
powdered stems (Khandelwal, 2005; Starmans & Nijhuis, 1996).
Defatting of Plant Material
62gram of roots of Aerva lanata Linn were coarsely powdered and subjected to extraction with
petroleum ether in maceration method. The extraction was continued till the defatting of the
material had taken place.
Extraction by macerationmethod
Defatted powdered roots of Aerva lanata Linn were exhaustively extracted with successive
solvent like chloroform, ethyl acetate, methanol and water by maceration method. The extract
was evaporated above their boiling points. Finally, the percentage yields were calculated of the
dried extracts (Mukherjee, 2007).
Phytochemical screening
Phytochemical examinations were carried out extracts as per the following standard methods
(Kokate, 1994: Pandey & Tripathi, 2014).
Separation and Identification of phytoconstituents by TLC
Thin layer chromatography is based on the adsorption phenomenon. In this type of
chromatography mobile phase containing the dissolved solutes passes over the surface of
stationary phase. Each solvent extract was subjected to thin layer chromatography (TLC) as
per conventional one dimensional ascending method using silica gel 60F254, 7X6 cm (Merck)
were cut with ordinary household scissors. Plate markings were made with soft pencil. Glass
capillaries were used to spot the sample for TLC applied sample volume 1-micro litre by using
capillary at distance of 1 cm at 5 tracks. In the twin trough chamber with different solvent
system toluene: ethyl acetate: formic acid (5:4:1) solvent system used for flavonoids and
toluene: ethyl acetate: formic acid (7:5:1) solvent system used for phenol (Patel et al., 2017).
After pre-saturation with mobile phase for 20 min for development were used. The movement
of the active compound was expressed by its retention factor (Rf), values were calculated for
different samples (Sherma & Fried, 2003).
Total phenol content estimation
The total phenol content of the extract was determined by the modified folin-ciocalteu method
(Parkhe and Bharti, 2019). 10 mg Gallic acid was dissolved in 10 ml methanol, various aliquots
of 10- 50µg/ml was prepared in methanol. 10 mg of dried extract was dissolved in 10 ml
methanol and filter. Two ml (1mg/ml) of this extract was for the estimation of phenol. 2 ml of
extract and each standard was mixed with 1 ml of Folin-Ciocalteu reagent (previously diluted
with distilled water 1:10 v/v) and 1 ml (7.5g/l) of sodium carbonate. The mixture was vortexed
for 15s and allowed to stand for 10min for colour development. The absorbance was measured
at 765 nm using a spectrophotometer.
Total flavonoids content estimation
Determination of total flavonoids content was based on aluminium chloride method (Parkhe
and Bharti, 2019).10 mg quercetin was dissolved in 10 ml methanol, and various aliquots of 5-

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1352
25μg/ml were prepared in methanol.10 mg of dried extract was dissolved in 10 ml methanol
and filter. Three ml (1mg/ml) of this extract was for the estimation of flavonoids.1 ml of 2%
AlCl3 solution was added to 3 ml of extract or each standard and allowed to stand for 15min at
room temperature; absorbance was measured at 420 nm.
Results & Discussion
The chloroform extract found to have percentage yield of 2.32% which is lowest of all. A Bit
good results are seen in case of ethyl acetate with percentage yield of 2.90%. The aqueous
extract posses 3.44% yield. The methanol extract has highest percentage yield of 8.14%. The
phytochemical screening of Aerva lanata was then performed to detect the phytochemical
constituents. The chloroform extract found to have presence of tannin only. In case of ethyl
acetate phenol & tannin tested positive. The methanolic extract yielded somewhat greater
number of phytoconstituents which are namely flavonoid, phenol & tannin. Finally, the
aqueous extract tested positive for flavonoid & tannin.
According to study conducted by Battu and Kumar the chloroform extract of the entire A.
lanata plant only tannins, alkaloids, and flavonoids in addition to carbohydrates and the lack
of alkaloids, proteins, saponins, and resins (Battuand Kumar,2012).
Similarly according to Yamunadeviet al. methanolic extract of A. lanata has reported the
presence of flavonoids and glycosides as in the current study, they have also shown the presence
of terpenoids and alkaloids (Yamunadeviet al., 2011).
After phytochemical screening Thin layer chromatography was performed. TLC for flavoinoid
was performed by using quercetin as flavonoid. The Rf value obtained for Quercetin standard
was 0.72. For methanolic extract of Aerva lanata, range of Rf values are obtained when
visualized in short UV. The Rf value obtained confirmed the compound present in extract are
part of large flavonoid groups. Additionally, TLC for phenol was performed for methanol, ethyl
acetate & aqueous extract. The Rf value obtained for gallic acid standard was 0.58. From the
Rf values obtained for all the three extract it was evident that ethyl acetate extract contain
phenol similar to gallic acid.
Total phenolic content for Aerva lanata methanolic extract estimated to be 1.380 mg/100mg
while the ethyl acetate & aqueous extract contain 0.866 & 0.613 mg/ 100 mg of phenol
respectively. Total flavonoid content was estimated only in methanolic extract which was
observed to be 1.280mg/ 100 mg.
In same way study conducted by Bahar et al., 2013 noticed that Aerva lanata extracts in
methanol and petroleum ether had total phenol concentrations of 108.9125 mg/ml and
147.5025 mg/ml, respectively.
Table No. 1: % yield of Aerva lanataLinn
S. No.
Extracts
% Yield (W/W)
1.
Chloroform
2.32%
2.
Ethyl acetate
2.90%
3.
Methanol
8.14%
4.
Aqueous
3.44%

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1353
Table No. 2: Result of phytochemical screening of Aerva lanata Linn
S.
No.
Constituents
Chloroformextract
Ethyl acetate
extract
Methanol
extract
Aqueous
extract
1.
Alkaloids
Hager’s Test:
-ve
-ve
-ve
-ve
2.
Glycosides
Conc. H2SO4 Test:
-ve
-ve
-ve
-ve
3.
Flavonoids
Lead acetate Test:
Alkaline Reagent
Test:
-ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
4.
Diterpenes
Copper acetate
Test:
-ve
-ve
-ve
-ve
5.
Phenol
Ferric Chloride
Test:
FolinCiocalteu
Test:
-ve
-ve
-ve
+ve
-ve
+ve
-ve
+ve
6.
Proteins
Xanthoproteic
Test:
-ve
-ve
-ve
-ve
7.
Carbohydrate
Fehling’s Test:
-ve
-ve
-ve
-ve
8.
Saponins
Froth Test:
-ve
-ve
-ve
-ve
9.
Tannins
Gelatin Test:
+ve
+ve
+ve
+ve
10.
Sterols
Salkowski’s Test:
-ve
-ve
-ve
-ve
[+ve =positive; -ve= negative]
Table No. 3:Identification of phytoconstituents by TLC of Aerva lanataLinn
TLC of Flavonoids
S.
No.
Mobile phase
Toluene: Ethyl acetate: Formic acid
(5:4:1)
Rfvalue
1.
(Quercetin)
Dis. travel by mobile phase= 5cm
No. of spot at long UV= 1
No. of spot at short UV = 1
Long- 0.72
Short- 0.72

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1354
No. of spot at normal light= 1
Normal- 0.72
2.
(Methanol extract)
Dis. travel by mobile phase= 5cm
No. of spot at long UV = 4
No. of spot at short UV = 5
No. of spot at normal light= 2
Long-0.1, 0.2, 0.8, 0.84
Short- 0.8, 0.86, 0.98, 0.9, 0.96
Normal- 0.8, 0.98
Spot Sequence
Quercetin
1st
Methanolic extract
2nd
Normal light Short UV Long UV
Figure 1: TLC of Flavonoids
Table No. 4: Identification of phytoconstituents by TLC of Aerva lanataLinn
TLC of Phenol
S.
No.
Mobile phase
Toluene: Ethyl acetate: Formic acid
(5:4:1)
Rfvalue
1.
(Gallic acid)
Dis. travel by mobile phase= 5cm
No. of spot at long UV= 1
No. of spot at short UV = 1
No. of spot at normal light= 1
Long- 0.58
Short- 0.58
Normal- 0.58

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1355
2.
(Methanol extract)
Dis. travel by mobile phase= 5cm
No. of spot at long UV = 1
No. of spot at short UV = 1
No. of spot at normal light= 1
Long-0.1
Short- 0.64
Normal- 0.74
3.
(Ethyl acetate extract)
Dis. travel by mobile phase= 5cm
No. of spot at long UV = 5
No. of spot at short UV = 5
No. of spot at normal light= 1
Long-0.58, 0.72, 0.76, 0.94, 0.98
Short- 0.1, 0.24, 0.64, 0.8, 0.82
Normal- 0.92
4.
(Aqueous extract)
Dis. travel by mobile phase= 5cm
No. of spot at long UV = 1
No. of spot at short UV = 5
No. of spot at normal light=1
Long- 0.42
Short- 0.42
Normal- 0.42
Spot Sequence
Gallic acid
1st
Methanolic extract
2nd
Ethyl acetate extract
3rd
Aqueous extract
4th
Normal light Short UV Long UV
Figure 2: TLC of phenol
Table No. 5: Estimation of total phenolic and flavonoids content of Aerva lanataLinn

Aerva lanata: Roots Extract for the analysis of phytochemicals
Eur. Chem. Bull. 2023,12(Special issue 12), 1349-1358 1356
S. No.
Extract
Total phenolic content
(mg/100mg of dried
extract)
Total flavonoids content
(mg/ 100 mg of dried
extract)
1
Ethyl acetate
0.866
-
2
Methanol
1.380
1.280
3
Aqueous
0.613
-
Conclusion
The traditional medical system does, however, provide physiologically active molecules that
are desirable sources of potential secondary metabolites that might be used as pharmaceuticals.
The objective of additional investigation is to identify the antibacterial and antioxidant
components that may be employed in herbal formulations. The results of this study provided a
crucial phytomarker for identifying and describing A. lanata. The structure of the bioactive
chemical will also be isolated, described, and clarified by pharmacological study.
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